Difference between revisions of "Part:BBa K258007"
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EGF-TliA, EGF-LARD1 fusion proteins were secreted when ABC transporter was present. E. coli harboring the PrtDEF of E. chrysanthemi secreted more fusion proteins than without ABC transporter. In fact, in enzyme activation assay(with Hitachi Spectrophotometer) we observed that although our protein -EGF- has disulfide bonds, it was excreted succesfully. In addition, when compearing with lack of ABC domain, we saw that protein excretion into medium (supernatant) is 10-fold more with ABC transporter-PrtDEF-. PrtDEF functioned well at 37°C, the optimum growth temperature for E. coli | EGF-TliA, EGF-LARD1 fusion proteins were secreted when ABC transporter was present. E. coli harboring the PrtDEF of E. chrysanthemi secreted more fusion proteins than without ABC transporter. In fact, in enzyme activation assay(with Hitachi Spectrophotometer) we observed that although our protein -EGF- has disulfide bonds, it was excreted succesfully. In addition, when compearing with lack of ABC domain, we saw that protein excretion into medium (supernatant) is 10-fold more with ABC transporter-PrtDEF-. PrtDEF functioned well at 37°C, the optimum growth temperature for E. coli | ||
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Sequence and Features PrtDEF: ABC transporter of Erwinia chrysanthemi for PrtA | Sequence and Features PrtDEF: ABC transporter of Erwinia chrysanthemi for PrtA | ||
EGF: Human Epidermal Growth Factor | EGF: Human Epidermal Growth Factor | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
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===Functional Parameters=== | ===Functional Parameters=== | ||
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Latest revision as of 00:39, 11 October 2009
Export of recombinant proteins in Escherichia coli using ABC transporter of Erwinia chrysanthemi
EGF-TliA, EGF-LARD1 fusion proteins were secreted when ABC transporter was present. E. coli harboring the PrtDEF of E. chrysanthemi secreted more fusion proteins than without ABC transporter. In fact, in enzyme activation assay(with Hitachi Spectrophotometer) we observed that although our protein -EGF- has disulfide bonds, it was excreted succesfully. In addition, when compearing with lack of ABC domain, we saw that protein excretion into medium (supernatant) is 10-fold more with ABC transporter-PrtDEF-. PrtDEF functioned well at 37°C, the optimum growth temperature for E. coli
Sequence and Features PrtDEF: ABC transporter of Erwinia chrysanthemi for PrtA EGF: Human Epidermal Growth Factor
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 210
Illegal PstI site found at 793
Illegal PstI site found at 1140
Illegal PstI site found at 3055 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 4344
Illegal PstI site found at 210
Illegal PstI site found at 793
Illegal PstI site found at 1140
Illegal PstI site found at 3055 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1259
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 210
Illegal PstI site found at 793
Illegal PstI site found at 1140
Illegal PstI site found at 3055 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 210
Illegal PstI site found at 793
Illegal PstI site found at 1140
Illegal PstI site found at 3055
Illegal NgoMIV site found at 4706
Illegal AgeI site found at 3189 - 1000COMPATIBLE WITH RFC[1000]